title: Deferoxamine promotes osteoblastic differentiation in human periodontal ligament cells via the nuclear factor erythroid 2-related factor-mediated antioxidant signaling pathway.
abstract: OBJECTIVE
  Recently it was reported that deferoxamine (DFO), an iron chelator, stimulates bone formation from MG63 and mesenchymal stem cells, but inhibits differentiation in rat calvarial cells; however, the effect of DFO on osteoblastic differentiation in human periodontal ligament cells (hPDLCs) has not been reported. The aim of this study was to investigate the effects and the possible underlying mechanism of DFO on osteoblastic differentiation of hPDLCs.
  
  
  METHODS
  The effect of DFO on osteoblast differentiation was determined by the staining intensity of calcium deposits with Alizarin red and by RT-PCR analysis of the expression of osteoblastic markers. Signal transduction pathways were analyzed by western blotting.
  
  
  RESULTS
  DFO increased osteogenic differentiation in a concentration-dependent manner by expression of the mRNA for differentiation markers and calcium nodule formation. Exposure of hPDLCs to DFO resulted in increases in the production of reactive oxygen species and in the levels of nuclear factor erythroid 2-related factor (Nrf2) protein in nuclear extractions, as well as a dose-dependent increase in the expression of Nrf2 target genes, including glutathione (GSH), glutathione S-transferase, γ-glutamylcysteine lygase, glutathione reductase and glutathione peroxidase. Pretreatment with Nrf2 small interfering RNA, GSH depletion by buthionine sulfoximine and diethyl maleate, and with antioxidants by N-acetylcysteine and vitamin E, blocked DFO-stimulated osteoblastic differentiation. Furthermore, pretreatment with GSH depletion and antioxidants blocked DFO-induced p38 MAPK, ERK, JNK and nuclear factor-kappaB pathways.
  
  
  CONCLUSIONS
  These data indicate, for the first time, that nontoxic DFO promotes osteoblastic differentiation of hPDLCs via modulation of the Nrf2-mediated antioxidant pathway.
authors.affiliation: Department of Periodontology, School of Dentistry, Kyung Hee University, Seoul, Korea.
authors.forename: J H
authors.initials: JH
authors.lastname: Chung
authors.forename: Y S
authors.initials: YS
authors.lastname: Kim
authors.forename: K
authors.initials: K
authors.lastname: Noh
authors.forename: Y M
authors.initials: YM
authors.lastname: Lee
authors.forename: S W
authors.initials: SW
authors.lastname: Chang
authors.forename: E C
authors.initials: EC
authors.lastname: Kim
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D061745:Antioxidant Response Elements / Q000187:drug effects*; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002113:Calcification, Physiologic / Q000187:drug effects; D002118:Calcium / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D003676:Deferoxamine / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016166:Free Radical Scavengers / Q000494:pharmacology; D005721:Glutamate-Cysteine Ligase / Q000032:analysis / Q000037:antagonists & inhibitors; D005978:Glutathione / Q000032:analysis / Q000037:antagonists & inhibitors; D005979:Glutathione Peroxidase / Q000032:analysis; D005980:Glutathione Reductase / Q000032:analysis; D005982:Glutathione Transferase / Q000032:analysis; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008298:Maleates / Q000494:pharmacology; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000494:pharmacology*; D016328:NF-kappa B / Q000187:drug effects; D010006:Osteoblasts / Q000187:drug effects*; D010513:Periodontal Ligament / Q000166:cytology* / Q000187:drug effects; D034741:RNA, Small Interfering / Q000235:genetics; D017382:Reactive Oxygen Species / Q000032:analysis; D017262:Siderophores / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D014810:Vitamin E / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; D008298:Maleates; D051267:NF-E2-Related Factor 2; D016328:NF-kappa B; C495635:NFE2L2 protein, human; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D017262:Siderophores; D014810:Vitamin E; D019328:Buthionine Sulfoximine; D005979:Glutathione Peroxidase; D005980:Glutathione Reductase; D005982:Glutathione Transferase; D005721:Glutamate-Cysteine Ligase; C014476:diethyl maleate; D005978:Glutathione; D003676:Deferoxamine; D002118:Calcium; D000111:Acetylcysteine
keywords: antioxidant; deferoxamine; iron chelator; nuclear factor erythroid 2-related factor; osteoblastic differentiation; periodontal ligament cells